Cat. No.
MAK-0044
Product Name
Rat Anti-DNP IgG
Storage
2-8℃
Species
Rat
Instructions
This kit is designed to measure anti-DNP IgG levels in serum collected 25 days after immunization with DNP-carrier protein conjugates, at which point the immune response originates primarily from IgG.
Introduction
Haptens such as dinitrophenol (DNP), when attached to carrier proteins such as ovalbumin or keyhole limpet hemocyanin (KLH) and injected into animals, produce a strong immune response. By measuring changes in the levels of anti-DNP IgM and IgG in appropriate animal models, researchers can assess the impact of pharmacologic or genetic manipulations on the immune system. Life Diagnostics, Inc. offers a simple ELISA for measurement of rat anti-DNP IgG. We also manufacture ELISA kits for rat anti-DNP IgM and mouse anti-DNP IgM and IgG.
Principle Of The Assay
The rat anti-DNP IgG test kit is based on a solid phase enzymelinked immunosorbent assay (ELISA). The assay uses DNP-BSA as the capture antigen (coated on the microtiter wells) and horseradish peroxidase (HRP) conjugated anti-rat IgG antibodies for detection. Serum or plasma samples are diluted and incubated
alongside standards in the microtiter wells for 45 minutes. The wells are subsequently washed, and HRP conjugate is added and incubated for 45 minutes. Anti-DNP IgG molecules are thus sandwiched between immobilized DNP and the detection antibody conjugate. The wells are then washed to remove unbound HRP
labeled antibodies, and TMB Reagent is added and incubated for 20 minutes at room temperature. This results in the development of a blue color. Color development is stopped by the addition of Stop Solution, changing the color to yellow, and optical density is measured spectrophotometrically at 450 nm. The concentration of anti-DNP IgG is proportional to the absorbance at 450 nm and is derived from a standard curve.
